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From Rarotonga, Cook Islands Biodiversity Data Journal 3: e4180 doi: 10.3897/BDJ.3.e4180 Taxonomic Paper Neoniphon pencei, a new species of holocentrid (Teleostei: Beryciformes) from Rarotonga, Cook Islands Joshua M. Copus†‡, Richard L. Pyle , John L. Earle‡ † Hawaii Institute of Marine Biology, Kaneohe, United States of America ‡ Bishop Museum, Honolulu, United States of America Corresponding author: Joshua M. Copus ([email protected]) Academic editor: Rupert Collins Received: 20 Oct 2014 | Accepted: 22 Jan 2015 | Published: 26 Jan 2015 Citation: Copus J, Pyle R, Earle J (2015) Neoniphon pencei, a new species of holocentrid (Teleostei: Beryciformes) from Rarotonga, Cook Islands. Biodiversity Data Journal 3: e4180. doi: 10.3897/BDJ.3.e4180 Abstract Neoniphon pencei, n. sp., is described from thirteen specimens, 132-197 mm standard length (SL) collected from mesophotic coral ecosystems (MCEs) at Rarotonga, Cook Islands by divers using mixed-gas closed-circuit rebreathers. It differs from all other species of the genus in number of lateral line scales, scales above and below lateral line, elements of life color, and in COI and cytochrome b DNA sequences. Of the five other known species of Neoniphon, it is most similar to the Indo-Pacific N. aurolineatus and the western Atlantic N. marianus both morphologically and genetically. Keywords Holocentridae, Neoniphon, new species, Mesophotic Coral Ecosystems, MCE, Rarotonga, Cook Islands © Copus J et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. 2 Copus J et al. Introduction Collections of shore fishes at mesophotic depths (~30-200 m) across the Indo-Pacific are yielding a surprising number of undescribed species (Pyle 2000). Here, we describe a new species of the genus Neoniphon from depths of 90-115 m at Rarotonga, Cook Islands, raising to six the number of species within this genus. The genus Neoniphon Castelnau 1875 includes five currently recognized species: N. argenteus (Valenciennes in Cuvier and Valenciennes 1831), N. aurolineatus (Liénard 1839), N. opercularis (Valenciennes in Cuvier and Valenciennes 1831) and N. sammara (Forsskål 1775) from the tropical Indo-Pacific, and N. marianus (Cuvier in Cuvier and Valenciennes 1829) from the tropical western Atlantic. Woods and Sonoda (1973) placed these five species (referring to N. argenteus by the junior synonym laevis [=læve] Günther 1859, and to N. aurolineatus by the junior synonym scythrops Jordan and Evermann 1903) within the genus Flammeo Jordan and Evermann 1898, on the grounds that the earlier name Neoniphon (type species: N. armatus Castelnau 1875) was "based on a species whose status is uncertain" (p. 345). Randall and Heemstra (1985) treated the four Indo- Pacific species of the genus, and confidently asserted that the original description of N. armatus Castelnau 1875 (the type species ofNeoniphon) is conspecific with N. sammara, and therefore considered Neoniphon as a valid genus with higher nomenclatural priority, to apply to the the four Indo-Pacific species as well as N. marianus from the tropical Atlantic. The lead author is working on a larger phylogeographic study involving this genus, which will be published at a later time. Materials and methods Type specimens of the new species Neoniphon pencei from Rarotonga, Cook Islands have been deposited in the Bernice P. Bishop Museum, Honolulu (BPBM); the California Academy of Sciences, San Francisco (CAS); and the U.S. National Museum of Natural History, Washington, D.C. (USNM). Measurements and counts given here follow the methods outlined in Randall (1998). Lengths of specimens are given as ratios of: standard length (SL) measured from the tip of the snout to the base of the caudal fin at the end of the hypural plate; body depth, taken at the point of maximum depth; or head length, measured from the median anterior point of the upper lip to the end of the longest opercular spine. Meristics and measurements were compared with data obtained from the literature for all five currently recognized species (Shimizu and Yamakawa 1979). Tissue samples were obtained from each of the thirteen individuals of N. pencei collected at Rarotonga, Cook Islands by spear at 90-115 m. Tissue samples were also obtained from twenty-two specimens of the five other species of Neoniphon: N. sammara (n=6) collected from Diego Garcia, British Indian Ocean Territory; N. opercularis (n=2) collected from Moorea, French Polynesia; N. aurolineatus (n=7) collected from Oahu, Hawaii; N. marianus Neoniphon pencei, a new species of holocentrid (Teleostei: Beryciformes) ... 3 (n=1) collected from the Commonwealth of the Bahamas; and N. argenteus (n=6) collected from the Republic of Kiritimati. Total genomic DNA was extracted from each sample using the 'HotSHOT' protocol (Meeker et al. 2007). A 577-bp fragment of the mtDNA cytochrome b ( Cyt b>) region was amplified using modified primers from Song et al. (1998) (5’- TGAAGTTGTCGGGATCTCCT-3’) and Taberlet et al. (1992) (5’- TGCCGTGACGTAAACTATGG-3’). Polymerase chain reaction (PCR) was performed in a 15 µl reaction containing 7.5 µl BioMix Red (Biolone Inc., Springfield, NJ, USA), 0.2 µM of each primer, 5-50 ng template DNA, and nanopure water (Thermo Scientific* Barnstead, Dubuque, IA, USA) to volume. PCR cycling parameters were as follows: initial 95°C denaturation for 10 min. followed by 35 cycles of 94°C for 30 sec, 60°C for 30 sec, and 72° C for 30 sec, followed by a final extension of 72°C for 10 min. PCR products were visualized using a 1.5% agarose gel with GelStarTM (Cambrex Bio Science Rockland, Inc., Rockland MA, USA) and then cleaned by incubating with 0.75 units of Exonuclease and 0.5 units of Shrimp Akaline Phosphate (ExoSAP; USB, Cleveland, OH, USA) per 7.5 µl of PCR product for 30 min. at 37°C followed by 85°C for 15 min. Sequencing was conducted in the forward direction and reverse direction when needed using a genetic analyzer (ABI 3130XL, Applied Biosystems, Foster City, California) at the Hawai'i Institute of Marine Biology EPSCoR Sequencing Facility. The sequences were aligned, edited and trimmed to a common length using Geneious Pro (v.5.6.6) DNA analysis software (Drummond et al. 2012). Twelve representative Cyt b> haplotypes were deposited in GenBank (accession numbers KJ188431-188436 and KJ201921-201926). jModelTest v.2.1.4 (Darriba et al. 2012, Guindon and Gascuel 2003) was used with an Akaike information criterion (AIC) test to determine the best nucleotide substitution model for the data. The GTR+G model with gamma parameter 0.1840 was identified to be the best suited model for phylogenetic inference. Maximum Likelihood, Neighbor-Joining, and Maximum Parsimony tree-building methods were implemented using Mega v.5.2.2 (Tamura et al. 2011). Sargocentron rubrum (Genbank accession number AP004432.1) was used to root a maximum likelihood phylogenetic reconstruction. Clade support was evaluated by bootstrapping 1,000 replicates in all cases (Felsenstein 1985). A DNA barcode (cytochrome c oxidase I; COI) was completed for the holotype and one paratype (BPBM 41196) using the primers from Baldwin et al. (2009), Fish-BCH (5'- ACTTCYGGGTGRCCRAARAATCA-3') and Fish-BCL (5'- TCAACYAATCAYAAAGATATYGGCAC-3') using the following PCR protocol: initial 95°C denaturation for 10 min. followed by 35 cycles of 94°C for 30 sec, 55°C for 30 sec, and 72° C for 30 sec, followed by a final extension of 72°C for 10 min. All other procedures were as described above. Both individuals possessed the same COI haplotype, so only one record was deposited in GenBank (http://www.ncbi.nlm.nih.gov/; accession number KJ188437) and BOLD (www.boldsystems.org; dx.doi.org/10.5883/DS-NPE511). 4 Copus J et al. Taxon treatment Neoniphon pencei Copus, Pyle, and Earle, sp. n. • ZooBank urn:lsid:zoobank.org:act:43F5CABA-6E4B-42BB-8569-8F93D3502DE9 • Barcode of Life NPE001-14 • GenBank KJ201926 • GenBank KJ188437 Materials Holotype: a. scientificName: Neoniphon pencei; originalNameUsage: Neoniphon pencei Copus, Pyle and Earle, 2014; originalNameUsageID: 43f5caba-6e4b-42bb-8569-8f93d3502de9; namePublishedIn: Copus, Joshua M., Richard L. Pyle & John L. Earle. 2014. Neoniphon pencei, a new species of holocentrid from Rarotonga, Cook Islands. Biodiversity Data Journal.; namePublishedInID: bbdce765-389b-4338-9c36-68def122f4fc; nameAccordingTo: Copus, Joshua M., Richard L. Pyle & John L. Earle. 2014. Neoniphon pencei, a new species of holocentrid from Rarotonga, Cook Islands. Biodiversity Data Journal.; nameAccordingToID: bbdce765-389b-4338-9c36-68def122f4fc; acceptedNameUsage: Neoniphon pencei Copus, Pyle and Earle, 2014; acceptedNameUsageID: 43f5caba-6e4b-42bb-8569-8f93d3502de9; taxonID: 43f5caba-6 e4b-42bb-8569-8f93d3502de9; scientificNameID: 43f5caba-6e4b-42bb-8569-8f93d3502d e9; parentNameUsageID: b047f156-f8da-4ec6-9f64-87345b68a759; parentNameUsage: Neoniphon Castelnau, 1875; higherClassification: Animalia; Deuterostomia; Chordata; Craniata; Gnathostomata; Actinopterygii; Beryciformes; Holocentroidei; Holocentridae; Neoniphon; kingdom: Animalia; phylum: Chordata; class: Actinopterygii; order: Beryciformes; family: Holocentridae; taxonRank: species; vernacularName: Pence's Squirrelfish; nomenclaturalCode: ICZN; genus: Neoniphon; specificEpithet: pencei; scientificNameAuthorship: Copus, Pyle & Earle; waterBody: Pacific Ocean; islandGroup: Cook Islands; island: Rarotonga; country: Cook Islands; countryCode: CK; locality: E side;
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